Quantitation of group-specific a antigen in hepatitis B vaccines by anti-HBs/a monoclonal antibody.
Balb/c mice were immunized with aluminium hydroxide [alum, Al (OH)3]-adjuvanted hepatitis B (HB) vaccines of subtypes adr, ayw or adw. Spleen cells from the immune animals were fused with SP2/O cells. Eight hybridoma clones producing antibodies specific for HB surface antigen (HBsAg) were selected. Monoclonal antibodies (mAbs) of four clones were specific for group-specific antigen/a, and the other of four clones were specific for subtype antigen/d, y, r, or w. The anti-HBs/a mAbs were classified into three non-competitive groups. Quantitation of group-specific determinant a of HBsAg (HBsAg/a) was performed by sandwich enzyme-linked immunosorbent assay (ELISA), in which a solid phase of anti-HBs guinea-pig polyclonal antibodies (pAb), the HBsAg for testing, anti-HBs/a mouse mAb and horseradish peroxidase (HRP)-conjugated anti-mouse IgG were used. The unadsorbed HBsAg was used to establish the standard curve of HBsAg/a. The lower detection limits were 0.5 to 1 ng/ml of HBsAg. Methods of solubilization of alum were investigated to quantify HBsAg/a in adsorbed HB vaccines. The recovery rate was more than 60% if vaccines were prediluted. The recovery of HBsAg/a in HB vaccines produced by the same manufacturer showed the similar recovery rate, and the contents of HBsAg/a in adsorbed HB vaccines could be estimated by the recovery rate determined for adsorbed HB vaccines.
Yamamoto
H
H
Department of Viral Disease and Vaccine Control, National Institute of Infectious Diseases, Tokyo, Japan.
Satoh
T
T
Kiyohara
T
T
Totsuka
A
A
Moritsugu
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D005260:Female; D006168:Guinea Pigs; D006510:Hepatitis B Antibodies / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000145:classification / Q000276:immunology*; D017325:Hepatitis B Vaccines / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines